SINGLE-DOSE ADMINISTRATION OF FACTOR VIIa

ABSTRACT

The present invention provides methods for preventing and/or treating bleeding episodes by administering a single dose of a Factor VIIa equivalent. Preferably, the single dose comprises between about 150 and about 500 ug/kg Factor VIIa equivalent.

CROSS REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. application Ser. No.15/634,153, filed Jun. 27, 2017, which is a continuation of U.S.application Ser. No. 15/249,598, filed Aug. 29, 2016 (now abandoned),which is a continuation of U.S. application Ser. No. 14/973,837, filedDec. 18, 2015 (now abandoned), which is a continuation of U.S.application Ser. No. 14/683,392, filed Apr. 10, 2015 (now abandoned),which is a continuation of U.S. application Ser. No. 12/127,917, filedMay 28, 2008 (now U.S. Pat. No. 9,029,324, issued May 12, 2015), whichis a continuation of U.S. application Ser. No. 10/196,902 filed Jul. 16,2002 (now U.S. Pat. No. 7,419,949, issued Sep. 2, 2008), and claimspriority under 35 U.S.C. 119 of U.S. application Ser. No. 60/305,720filed on Jul. 16, 2001; the contents of which are fully incorporatedherein by reference.

SEQUENCE LISTING

The instant application contains a Sequence Listing which has beensubmitted in ASCII format via EFS-Web and is hereby incorporated byreference in its entirety. Said ASCII copy, created on Feb. 6, 2018, isnamed 6361US05 SeqList.txt and is 5 kilobytes in size.

FIELD OF THE INVENTION

The present invention relates to methods for preventing and/or treatingbleeding using coagulation factors.

BACKGROUND OF THE INVENTION

Factor VII is a plasma coagulation factor, which, once activated toFactor VIIa, initiates the normal haemostatic process by forming acomplex with tissue factor (TF), a cell surface glycoprotein that isexposed to the circulation as a result of injury to the vessel wall.Subsequently, the Factor VIIa-TF complex activates Factor IX and FactorX into their activated forms (Factor IXa and Factor Xa, respectively).Factor Xa converts limited amounts of prothrombin to thrombin on thetissue factor-bearing cell. Thrombin activates platelets and Factors Vand VIII into Factors Va and VIIIa, both cofactors in the furtherprocess leading to the full thrombin burst. Thrombin finally convertsfibrinogen to fibrin resulting in formation of a fibrin clot. Fibrinclots formed in the presence of high thrombin concentrations comprise atighter network and are more resistant to proteolysis than clots formedin lower concentrations of thrombin. Accordingly, a full thrombin burstis likely to be important for forming a hemostatic plug that isresistant to fibrinolysis and thus to facilitate full hemostasis andwound healing.

Factor VIIa, as well as Factor VIII and Factor IX, have been used tocontrol bleeding disorders that are caused by clotting factordeficiencies (such as, e.g. haemophilia A and B or deficiency ofcoagulation Factors XI or VII) or clotting factor inhibitors. FactorVIIa has also been used to control excessive bleeding caused bydefective platelet function, thrombocytopenia or von Willebrand'sdisease.

Typically, however, patients are treated with multiple injections orinfusions of a coagulation factor before the bleeding is stopped. In thecase of Factor VIII and Factor IX administration, a considerable numberof injections are needed to maintain haemostasis until the injurycausing the bleeding is completely healed. A quicker and more effectivetreatment, as well as a reduction in the number of injections neededbefore the bleeding is stopped, represent important benefit to suchpatients. It would also be a considerable benefit to a patient needingfrequent injections or infusions with a haemostatic agent that theinjection frequency be reduced.

Thus, there is a need in the art for methods for preventing and/ortreating bleeding episodes that reduce the duration of administrationand provide a more rapid hemostasis.

SUMMARY OF THE INVENTION

The present invention relates to methods for preventing and/or treatinga bleeding episode in a subject in need of such treatment, which arecarried out by administering to the subject, in a single dose, asingle-dose-effective amount of Factor VIIa or a Factor VIIa equivalent.Preferably, subsequent to the administration, no further Factor VIIa orprotein having Factor VIIa coagulant activity is administered to thesubject for an interval of at least about 1 hour. In some embodiments,the interval is at least about 4 hours; in other embodiments, theinterval is at least about 24 hours; and in some embodiments, no furtherFactor VIIa or protein having Factor VIIa coagulant activity isadministered during the particular bleeding episode that is beingtreated.

In some embodiments, the single-dose-effective amount comprises betweenabout 150 and about 500 ug/kg Factor VIIa or a corresponding amount of aFactor VIIa equivalent; in other embodiments, the single-dose-effectiveamount comprises between about 200 and about 500 ug/kg; between about250 and about 500 ug/kg; between about 300 and about 500 ug/kg; betweenabout 350 and 500 ug/kg; between about 400 and about 500 ug/kg; betweenabout 450 and about 500 ug/kg; and greater than 500 ug/kg, respectively,of Factor VIIa or a corresponding amount of a Factor VIIa equivalent.

In some embodiments, the Factor VIIa equivalent exhibits at least about30% of the coagulant activity of Factor VIIa on a molar basis.Non-limiting examples of a Factor VIIa equivalent include S52A-FVII,S60A-FVII; L305V-FVII, L305V/M306D/D309S-FVII, L305I-FVII, L305T-FVII,F374P-FVII, V158T/M298Q-FVII, V158D/E296V/M298Q-FVII, K337A-FVII,M298Q-FVII, V158D/M298Q-FVII, L305V/K337A-FVII,V158D/E296V/M298Q/L305V-FVII, V158D/E296V/M298Q/K337A-FVII,V158D/E296V/M298Q/L305V/K337A-FVII, K157A-FVII, E296V-FVII,E296V/M298Q-FVII, V158D/E296V-FVII, V158D/M298K-FVII, and S336G-FVII;Factor VIIa variants exhibiting increased proteolytic stability asdisclosed in U.S. Pat. No. 5,580,560; Factor VIIa that has beenproteolytically cleaved between residues 290 and 291 or between residues315 and 316; oxidized forms of Factor VIIa; Factor VII-sequence variantswherein the amino acid residue in positions 290 and/or 291 (of SEQ IDNO:1), preferably 290, have been replaced, and Factor VII-sequencevariants wherein the amino acid residue in positions 315 and/or 316 (ofSEQ ID NO:1), preferably 315, have been replaced.

In some embodiments, the method further comprises administering, with orsubstantially simultaneously with the single dose, a second coagulantagent. Non-limiting examples of a second coagulant agent include FactorVIII, Factor IX, and Factor XIII.

In some embodiments, the invention provides a method for treating ableeding episode, which is carried out by administering to a subject inneed of such treatment (i) a first amount of Factor VIIa or a FactorVIIa equivalent and (ii) a second amount of second coagulant agent,wherein the first and second amounts together comprise an aggregateeffective amount for treating the bleeding episode and the aggregateeffective amount is administered in a single dose.

In some embodiments, the Factor VIIa used in practicing the invention isrecombinant human Factor VIIa.

In one embodiment, the invention provides a method for treating ableeding episode, which is carried out by administering to a humansubject in need of such treatment an effective amount for treating saidbleeding of human Factor VIIa or a human Factor VIIa equivalent,wherein:

(i) said effective amount is administered in a single dose over a periodof less than about 5 minutes;

(ii) said effective amount comprises between about 300 and about 500ug/kg human Factor VIIa or human Factor VIIa equivalent or acorresponding amount of a Factor VIIa equivalent; and

(iii) subsequent to said administration, no further Factor VIIa orFactor VIIa equivalent is administered to said subject for a period ofat least about 1 hour.

In practicing the present invention, administration may be achieved byany mode of administration, including, without limitation, intravenous,intramuscular, subcutaneous, mucosal, and pulmonary routes ofadministration.

In another aspect, the invention provides a method for preventing ableeding episode, which is carried out by administering to a humansubject in need of such prevention an effective amount for preventingthe bleeding episode of human Factor VIIa or a human Factor VIIaequivalent, wherein:

(i) the effective amount is administered in a single dose over a periodof less than about 5 minutes;

(ii) the effective amount comprises between about 250 and about 500ug/kg human Factor VIIa or human Factor VIIa equivalent or acorresponding amount of a Factor VIIa equivalent; and

(iii) subsequent to the administration, no further Factor VIIa or FactorVIIa equivalent is administered to said subject for a period of at leastabout 1 hour.

In some embodiments, the subject suffers from hemophilia A or B.

In some embodiments, the bleeding is joint bleeding.

In some embodiments, the subject has not been treated therapeuticallywith an anticoagulant for at least about 48 hours prior toadministration of Factor VIIa or a Factor VIIa equivalent.

In some embodiments, the subject has not been treated therapeuticallywith a Vitamin K antagonist for at least about 48 hours prior to saidadministering.

DETAILED DESCRIPTION OF THE INVENTION

The present invention relates to methods for preventing and/or treatingbleeding in animals, particularly in humans. The invention is based onthe discovery that administration of a single dose comprising apredetermined amount of Factor VIIa is effective in treating bleedingepisodes, including major bleeding episodes, and can also be used toprevent an anticipated bleeding episode.

Without wishing to be bound by theory, it is believed that Factor VIIaenhances thrombin generation on tissue factor-bearing cells and onactivated platelets at the site of injury and that administration of asingle dose of Factor VIIa according to the present invention provides afull thrombin burst, thereby facilitating the formation of a tight,strong fibrin network that is relatively resistant to prematurefibrinolysis and obviating the need for further administration of FactorVIIa.

As used herein, prevention refers to prophylactic administration ofFactor VIIa so as to minimize or inhibit an anticipated bleedingepisode, such as, e.g., prior to surgery. Treatment refers to regulationof an already-occurring bleeding, such as, for example, in trauma, withthe purpose of inhibiting or minimizing the bleeding. It will beunderstood that efficacy in prevention and/or treatment according to thepresent invention encompasses the absence of significant side-effects,including, without limitation, disseminated intravascular coagulation(DIC), that would counterindicate to those of ordinary skill in the artthe use of any particular therapeutic regimen.

Bleeding refers to extravasation of blood from any component of thecirculatory system. The term “bleeding episode” includes, withoutlimitation, bleeding (including, without limitation, excessive,uncontrolled bleeding, i.e., haemorrhaging) in connection with surgeryor trauma, such as, for example, in connection with acute haemarthroses(bleedings in joints), chronic haemophilic arthropathy, haematomas,(e.g., muscular, retroperitoneal, sublingual and retropharyngeal),bleedings in other tissue, haematuria (bleeding from the renal tract),cerebral haemorrhage, surgery (e.g., hepatectomy), dental extraction,and gastrointestinal bleedings (e.g., UGI bleeds). Also included arehemorrhages in organs such as the brain, inner ear region and eyes withlimited possibility for surgical haemostasis; as well as hemorrhages inconnection with biopsies of various organs (liver, lung, tumour tissue,gastrointestinal tract) as well as laparoscopic surgery. Common to thesesituations is the difficulty in providing haemostasis using surgicaltechniques (such as, e.g., sutures or clips), which is also the casewhen bleeding is diffuse.

In practicing the invention, Factor VIIa or a Factor VIIa equivalent isadministered to a patient as a single dose comprising asingle-dose-effective amount. Administration of a single dose refers toadministration of an entire dose of Factor VIIa as a bolus over a periodof less than about 5 minutes. In some embodiments, the administrationoccurs over a period of less than about 2.5 minutes, and, in some, overless than about 1 min.

A single-dose-effective amount of Factor VIIa or a Factor VIIaequivalent refers to the amount of Factor VIIa or equivalent which, whenadministered in a single dose according to the invention, produces ameasurable improvement in at least one clinical parameter of haemostasisknown to those of ordinary skill in the art (see below). Typically, asingle-dose effective amount comprises at least 150 ug/kg Factor VIIa.In different embodiments, a single-dose-effective amount of Factor VIIacomprises between about 150-500 ug/kg; 250-500 ug/kg; 300-500 ug/kg;350-500 ug/kg; 400-500 ug/kg; 450-500 ug/kg; or more than 500 ug/kg,respectively. When Factor VIIa equivalents are administered according tothe present invention, a single-dose effective amount corresponding tothe above-cited amounts may be determined by comparing the anticoagulantactivity of the Factor VIIa equivalent with that of Factor VIIa (seebelow) and adjusting the amount to be administered proportionately.

It will be understood that a single-dose-effective amount of Factor VIIamay vary according to the subject's haemostatic status, which, in turn,may be reflected in one or more clinical parameters, including, e.g.,relative levels of circulating coagulation factors; amount of bloodlost; rate of bleeding; hematocrit, and the like. It will be furtherunderstood that the single-dose-effective amount may be determined bythose of ordinary skill in the art by routine experimentation, byconstructing a matrix of values and testing different points in thematrix.

In some embodiments, following administration of a single-dose of FactorVIIa or a Factor VIIa equivalent according to the invention, the patientreceives no further Factor VIIa or Factor VIIa equivalent for aninterval of at least about 1 hour. In some embodiments thepost-administration interval is at least about 4 hours; in otherembodiments, the post-administration interval is at least about 24hours. In still other embodiments, no further Factor VIIa or Factor VIIaequivalent is administered to treat the particular bleeding episode.

According to the invention, Factor VIIa or a Factor VIIa equivalent maybe administered by any effective route, including, without limitation,intravenous, intramuscular, subcutaneous, mucosal, and pulmonary routesof administration. Preferably, administration is by an intravenousroute.

Factor VIIa and Factor VIIa equivalents: In practicing the presentinvention, any Factor VIIa or equivalent may be used that is effectivein preventing or treating bleeding when administered in a single dose.In some embodiments, the Factor VIIa is human Factor VIIa, as disclosed,e.g., in U.S. Pat. No. 4,784,950 (wild-type Factor VII). The term“Factor VII” is intended to encompass Factor VII polypeptides in theiruncleaved (zymogen) form, as well as those that have beenproteolytically processed to yield their respective bioactive forms,which may be designated Factor VIIa. Typically, Factor VII is cleavedbetween residues 152 and 153 to yield Factor VIIa.

Factor VIIa equivalents include, without limitation, Factor VIIpolypeptides that have either been chemically modified relative to humanFactor VIIa and/or contain one or more amino acid sequence alterationsrelative to human Factor VIIa. Such equivalents may exhibit differentproperties relative to human Factor VIIa, including stability,phospholipid binding, altered specific activity, and the like.

In one series of embodiments, a Factor VIIa equivalent includespolypeptides that exhibit at least about 10%, preferably at least about30%, more preferably at least about 50%, and most preferably at leastabout 70%, of the specific biological activity of human Factor VIIa. Forpurposes of the invention, Factor VIIa biological activity may bequantified by measuring the ability of a preparation to promote bloodclotting using Factor VII-deficient plasma and thromboplastin, asdescribed, e.g., in U.S. Pat. No. 5,997,864. In this assay, biologicalactivity is expressed as the reduction in clotting time relative to acontrol sample and is converted to

“Factor VII units” by comparison with a pooled human serum standardcontaining 1 unit/ml Factor VII activity. Alternatively, Factor VIIabiological activity may be quantified by (i) measuring the ability ofFactor VIIa or a Factor VIIa equivalent to produce of Factor Xa in asystem comprising TF embedded in a lipid membrane and Factor X. (Perssonet al., J. Biol. Chem. 272: 19919-19924, 1997); (ii) measuring Factor Xhydrolysis in an aqueous system (see, Example 5 below); (iii) measuringthe physical binding of Factor VIIa or a Factor VIIa equivalent to TFusing an instrument based on surface plasmon resonance (Persson, FEBSLetts. 413:359-363, 1997) and (iv) measuring hydrolysis of a syntheticsubstrate by Factor VIIa and/or a Factor VIIa equivalent.

Non-limiting examples of Factor VII-related polypeptides havingsubstantially the same or improved biological activity as wild-typeFactor VII include S52A-FVII, S60A-FVII (lino et al., Arch. Biochem.Biophys. 352: 182-192, 1998); L305V-FVII, L305V/M306D/D309S-FVII,L305I-FVII, L305T-FVII, F374P-FVII, V158T/M298Q-FVII,V158D/E296V/M298Q-FVII, K337A-FVII, M298Q-FVII, V158D/M298Q-FVII,L305V/K337A-FVII, V158D/E296V/M298Q/L305V-FVII,V158D/E296V/M298Q/K337A-FVII, V158D/E296V/M298Q/L305V/K337A-FVII,K157A-FVII, E296V-FVII, E296V/M298Q-FVII, V158D/E296V-FVII,V158D/M298K-FVII, and S336G-FVII; Factor VIIa variants exhibitingincreased proteolytic stability as disclosed in U.S. Pat. No. 5,580,560;Factor VIIa that has been proteolytically cleaved between residues 290and 291 or between residues 315 and 316 (Mollerup et al., Biotechnol.Bioeng. 48: 501-505, 1995); oxidized forms of Factor VIIa (Kornfelt etal., Arch. Biochem. Biophys. 363: 43-54, 1999), Factor VII-sequencevariants wherein the amino acid residue in positions 290 and/or 291 (ofSEQ ID NO:1), preferably 290, have been replaced, and FactorVII-sequence variants wherein the amino acid residue in positions 315and/or 316 (of SEQ ID NO:1), preferably 315, have been replaced.

Preparations and formulations: The present invention encompassestherapeutic administration of Factor VIIa or Factor VIIa equivalents,which is achieved using formulations that comprise Factor VIIapreparations. As used herein, a “Factor VII preparation” refers to aplurality of Factor VIIa polypeptides or Factor VIIa equivalentpolypeptides, including variants and chemically modified forms, thathave been separated from the cell in which they were synthesized,whether a cell of origin or a recombinant cell that has been programmedto synthesize Factor VIIa or a Factor VIIa equivalent.

Separation of polypeptides from their cell of origin may be achieved byany method known in the art, including, without limitation, removal ofcell culture medium containing the desired product from an adherent cellculture; centrifugation or filtration to remove non-adherent cells; andthe like.

Optionally, Factor VII polypeptides may be further purified.Purification may be achieved using any method known in the art,including, without limitation, affinity chromatography, such as, e.g.,on an anti-Factor VII antibody column (see, e.g., Wakabayashi et al., J.Biol. Chem. 261: 11097, 1986; and Thim et al., Biochem. 27: 7785, 1988);hydrophobic interaction chromatography; ion-exchange chromatography;size exclusion chromatography;

electrophoretic procedures (e.g., preparative isoelectric focusing(IEF), differential solubility (e.g., ammonium sulfate precipitation),or extraction and the like. See, generally, Scopes, ProteinPurification, Springer-Verlag, New York, 1982; and Protein Purification,J.-C. Janson and Lars Ryden, editors, VCH Publishers, New York, 1989.Following purification, the preparation preferably contains less thanabout 10% by weight, more preferably less than about 5% and mostpreferably less than about 1%, of non-Factor VII proteins derived fromthe host cell.

Factor VII and Factor VII-related polypeptides may be activated byproteolytic cleavage, using Factor XIIa or other proteases havingtrypsin-like specificity, such as, e.g., Factor IXa, kallikrein, FactorXa, and thrombin. See, e.g., Osterud et al., Biochem. 11: 2853 (1972);Thomas, U.S. Pat. No. 4,456,591; and Hedner et al., J. Clin. Invest. 71:1836 (1983). Alternatively, Factor VII may be activated by passing itthrough an ion-exchange chromatography column, such as Mono Q®(Pharmacia) or the like. The resulting activated Factor VII may then beformulated and administered as described below. Pharmaceuticalcompositions or formulations for use in the present invention comprise a

Factor VIIa preparation in combination with, preferably dissolved in, apharmaceutically acceptable carrier, preferably an aqueous carrier ordiluent. A variety of aqueous carriers may be used, such as water,buffered water, 0.4% saline, 0.3% glycine and the like. The preparationsof the invention can also be formulated into liposome preparations fordelivery or targeting to the sites of injury. Liposome preparations aregenerally described in, e.g., U.S. Pat. Nos. 4,837,028, 4,501,728, and4,975,282. The compositions may be sterilised by conventional,well-known sterilisation techniques. The resulting aqueous solutions maybe packaged for use or filtered under aseptic conditions andlyophilised, the lyophilised preparation being combined with a sterileaqueous solution prior to administration.

The compositions may contain pharmaceutically acceptable auxiliarysubstances or adjuvants, including, without limitation, pH adjusting andbuffering agents and/or tonicity adjusting agents, such as, for example,sodium acetate, sodium lactate, sodium chloride, potassium chloride,calcium chloride, etc.

Combinations: The present invention encompasses combined single-doseadministration of an additional agent in concert with Factor VIIa or aFactor VIIa equivalent. In some embodiments, the additional agentcomprises a coagulant, including, without limitation, a coagulationfactor such as, e.g., Factor VIII, Factor IX, or Factor XIII; or aninhibitor of the fibrinolytic system, such as, e.g., aprotinin, c-aminocaproic acid or tranexamic acid. In other embodiments, theadditional agent comprises an anticoagulant, including, withoutlimitation, heparin, warfarin, coumarin, and modified Factor VIIpolypeptides, such as, e.g., R152E-Factor VIIa (Wildgoose et al.,Biochem 29: 3413-3420, 1990), S344A-Factor VIIa (Kazama et al., J. Biol.Chem. 270: 66-72, 1995), FFR-Factor VIIa (Hoist et al., Eur. J. Vasc.Endovasc. Surg. 15: 515-520, 1998), Factor VIIa lacking the Gla domain,(Nicolaisen et al., FEBS Letts. 317: 245-249, 1993), and chemicallymodified Factor VII polypeptides (U.S. Pat. No. 5,997,864).

It will be understood that, in embodiments comprising single-doseadministration of combinations of Factor VIIa with other agents, thedosage of Factor VIIa or Factor VIIa equivalent may on its own comprisea single-dose-effective amount. Alternatively, the combination of FactorVIIa or equivalent and the second agent may together comprise asingle-dose-effect amount for preventing or treating bleeding episodes.

Indications: The present invention encompasses single-doseadministration of Factor VIIa or a Factor VIIa equivalent to any patientwho either anticipates a bleeding episode or who is activelyexperiencing a bleeding episode. Such patients include, withoutlimitation, those suffering from bleeding disorders that are caused byclotting factor deficiencies (e.g. haemophilia A and B, or deficiency ofcoagulation Factors XI or VII); clotting factor inhibitors; defectiveplatelet function; thrombocytopenia; or von Willebrand's disease. Themethods of the invention may also be applied to patients who are aboutto undergo surgery, preferably major surgery, whether or not they sufferfrom a bleeding disorder; as well as trauma patients. Furthermore, anytype of profuse bleeding from the gastrointestinal tract, or anybleeding occurring postoperatively (including that occurring in patientsnot suffering from a bleeding disorder) may benefit from treatmentaccording to the present invention.

In some embodiments, the invention does not encompass administration ofFactor VIIa or equivalent to patients undergoing minor surgery. In otherembodiments, the invention does not encompass administration of FactorVIIa or equivalent to patients not suffering from a clotting disorderwho had been administered an anticoagulant (such as, e.g.,acetocoumerol) within 48 hours prior to Factor VIIa administration. Inother embodiments, the invention does not encompass administration ofFactor VIIa or equivalent to patients not suffering from a clottingdisorder who had been administered a Vitamin K antagonist within 48hours prior to Factor VIIa administration.

The present invention also provides the benefit of allowing a patient toself-administer an effective dose of Factor VIIa or a Factor VIIaequivalent in order to facilitate effective management of anticipated orcurrent bleeding episodes.

Many variations of the present invention will suggest themselves tothose skilled in the art in light of the above detailed description.Such obvious variations are within the full intended scope of theinvention.

All patents, patent applications, and literature references referred toherein are hereby incorporated by reference in their entirety.

The following examples are intended as non-limiting illustrations of thepresent invention.

EXAMPLE 1 High-Dose Factor VIIa Administration

Patients suffering from hemophilia (including, e.g., patients withclotting factor inhibitors, acquired inhibitor patients, patientssuffering from Factor VII deficiency, and patients suffering from vonWillebrands disease) are enrolled in a registry that tracks the outcomeof bleeding episodes whose treatment includes bolus administration ofFactor VIIa.

The bleeding episodes are characterized as spontaneous, related totraumatic injury, or other (including, e.g., surgical or dentalprocedures). The dosage groups are characterized as <100 ug/kg; 100-150ug/kg; 150-200 ug/kg; and >200 ug/kg. Responses to treatment (asassessed at 72 h) are characterized as cessation of bleeding, slowing ofbleeding; or no response.

EXAMPLE 2 Single High-Dose Factor VIIa Administration Description ofClinical Trial:

A randomized, multicenter, cross-over, double-blind study is performedto evaluate the efficacy and safety of Factor VIIa (by two differentblinded dose schedules) in producing hemostasis in joint bleeds in ahome-treatment setting. Subjects with congenital hemophilia A or B andinhibitors to Factor VIII or Factor IX receive treatment and areassessed for at least 9 hours after the dosing. The success or failureof the treatment is ascertained using a pilot algorithm to assesschanges in pain and joint mobility. rFVIIa will be given as anintravenous bolus injection, either at 270 μg/kg body weight dose athour 0, or at 90 μg/kg body weight doses given at hours 0, 3 and 6 andplacebo solutions will be administered to blind subject as to the doseregimen of rFVIIa being administered. If additional doses of Factor VIIaare administered within the first 9 hours to achieve hemostasis, thenthe treatment efficacy is graded as a failure.

Trial Population:

Twenty-four patients with congenital hemophilia A or B and inhibitors tofactor VIII or IX have been enrolled in this trial. Patients must haveexperienced two or more mild or moderate joint bleeds during the past 12months.

Assessments

Treatment efficacy is based on the evaluation of pain, joint mobilityand measure of circumference of the elbow or knee at the midpoint of thejoint in extension. These variables are graded and entered in the diaryby the patient/caregiver. Pain and mobility are assessed as more, nodifference or less than before the treatment and circumference ismeasured in millimeters. An independent blinded committee reviews thediary data on pain and joint mobility and judges the response totreatment as success or failure based on a pilot algorithm developed forthis study.

1. A method for treating a bleeding episode in a subject in need of suchtreatment, said method comprising administering intravenously to saidsubject a Factor VIIa equivalent, wherein said administering is in asingle dose and said dose comprises a single-dose-effective amount ofsaid Factor VIIa equivalent and wherein, subsequent to saidadministration, no further Factor VIIa equivalent is administered tosaid subject for a period of at least 4 hours.
 2. A method as defined inclaim 1, wherein said period is at least about 24 hours.
 3. A method asdefined in claim 1, wherein said single dose is administered over aperiod of less than about 5 minutes.
 4. A method as defined in claim 1,wherein said single-dose-effective amount comprises between about 300and about 500 ug/kg Factor VIIa equivalent.
 5. A method as defined inclaim 1, wherein said Factor VIIa equivalent exhibits at least about 30%of the coagulant activity of Factor VIIa on a molar basis.
 6. A methodas defined in claim 1, wherein said Factor VIIa equivalent is selectedfrom the group consisting of: S52A-FVII, S60A-FVII; L305V-FVII,L305V/M306D/D309S-FVII, L305I-FVII, L305T-FVII, F374P-FVII,V158T/M298Q-FVII, V158D/E296V/M298Q-FVII, K337A-FVII, M298Q-FVII, V158D/M298Q-FVII, L305V/K337A-FVII, V158D/E296V/M298Q/L305V-FVII,V158D/E296V/M298Q/K337A-FVII, V158D/E296V/M298Q/L305V/K337A-FVII,K157A-FVII, E296V-FVII, E296V/M298Q-FVII, V158D/E296V-FVII,V158D/M298K-FVII, and S336G-FVII; Factor VIIa that has beenproteolytically cleaved between residues 290 and 291; Factor VIIa thathas been proteolytically cleaved between residues 315 and 316; andoxidized forms of Factor VIIa.
 7. A method as defined in claim 1,further comprising administering, in said single dose, or substantiallysimultaneously with said single dose, a second coagulant agent.
 8. Amethod as defined in claim 7, wherein said second coagulant agent isselected from the group consisting of Factor VII, Factor VIII, FactorIX, and Factor XIII
 9. A method as defined in claim 1, furthercomprising administering an anticoagulant, wherein said anticoagulant isadministered in said single dose, or substantially simultaneously withsaid single dose.
 10. A method for treating a bleeding episode, saidmethod comprising administering to a subject in need of such treatment(i) a first amount of a Factor VIIa equivalent and (ii) a second amountof second coagulant agent, wherein said first and second amountstogether comprise an aggregate effective amount for treating saidbleeding and said aggregate effective amount is administered in a singledose.
 11. A method as defined in claim 1, wherein said administering isvia a route selected from the group consisting of intravenous,intramuscular, subcutaneous, mucosal, and pulmonary administration.